Engineering Cytochrome P450BM3 Enzymes for Direct Nitration of Unsaturated Hydrocarbons.

Applications of the peroxidase activity of cytochrome P450 enzymes in synthetic chemistry remain largely unexplored. We present herein a protein engineering strategy to increase cytochrome P450BM3 peroxidase activity for the direct nitration of aromatic compounds and terminal aryl-substituted olefins in the presence of a dual-functional small molecule (DFSM). Site-directed mutations of key active-site residues allowed the efficient regulation of steric effects to limit substrate access and, thus, a significant decrease in monooxygenation activity and increase in peroxidase activity.

Enabling highly ()-enantioselective epoxidation of styrene by engineering unique non-natural P450 peroxygenases.

Unlike the excellent ()-enantioselective epoxidation of styrene performed by natural styrene monooxygenases (ee > 99%), the ()-enantioselective epoxidation of styrene has not yet achieved a comparable efficiency using natural or engineered oxidative enzymes. This report describes the HO-dependent ()-enantioselective epoxidation of unfunctionalized styrene and its derivatives by site-mutated variants of a unique non-natural P450BM3 peroxygenase, working in tandem with a dual-functional small molecule (DFSM). The observed ()-enantiomeric excess (ee) of styrene epoxidation is up to 99% with a turnover number (TON) of 918 by the best enantioselective mutant F87A/T268I/L181Q, while the best active mutant F87A/T268I/V78A/A184L (with 98% ee) gave a catalytic TON of 4350, representing the best activity of a P450 peroxygenase towards styrene epoxidation to date.

Targeting of Rho kinase ameliorates impairment of diabetic endothelial function in intrarenal artery.

Endothelial dysfunction in kidney vasculature is the initial and key element for nephropathy in diabetes mellitus. Accumulating evidence suggests the protective role of Rho kinase inhibitors in endothelial dysfunction via modulating eNOS activity and NO production. However, the role of Rho kinase in diabetes-related endothelial dysfunction in kidney vasculature and the relevant mechanisms remain unknown.

Effect of treadmill training with body weight support on gait and gross motor function in children with spastic cerebral palsy.

Objective: To examine the effect of treadmill training with body weight support (TBWS) on gait and gross motor function in children with spastic cerebral palsy (CP).

Design: Eight children with spastic CP participated in the study. Their temporal-distance gait parameters, Gross Motor Function Measure, muscle tone, and selective motor control were assessed three times: two times under their regular therapeutic treatment (condition A), and one time after receiving the TBWS treatment in addition to their regular therapeutic treatments (condition B).

[Protective effects of puerarin on radiation injury of experimental rats].

Objective: To observe the protective effects of puerarin on radiation injury of experimental rats and to discuss the possible mechanism of its radiation protection.

Methods: Wistar rats were divided randomly into 4 groups with 8 rats in each group: physiological saline non-radiation (SN) group, puerarin non-radiation (PN) group, physiological saline radiation (SR) group, puerarin radiation (PR) group. The source of radiation was cobalt-60 gamma rays, and the rats were exposed to radiation (1.